Proteoglycan regulation of tumor angiogenesis and endothelial cell autophagy

蛋白多糖对肿瘤血管生成和内皮细胞自噬的调节

• 批准号: 10634656
• 负责人: RENATO V. IOZZO
• 金额: $ 37.79万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10634656
• 关键词: Affect; Affinity; Allografting; Angiogenesis Inhibitors; Angiogenic Factor; Angiostatic Proteins; Autophagocytosis; Binding; Biological; Biology; Breast Cancer cell line; Breast Carcinoma; C-terminal; Cancer Biology; Cells; Cellular Stress; Chemicals; Communication; Complex; Core Protein; Cues; Development; Disease; EIF-2alpha; Eating; Endothelial Cells; Endothelium; Enzymes; Exons; Female; Fibroblast Growth Factor Receptors; Funding; Gene Targeting; Genetic; Genetic Transcription; Grant; Growth; HAS2 gene; Heparan Sulfate Proteoglycan; Hyaluronan; Immunologics; In Vitro; Integrins; KDR gene; Knowledge; Laboratories; Ligation; Link; Malignant Neoplasms; Mediating; Methodology; Modality; Modeling; Molecular; Mus; Mutate; Names; Neoplasm Metastasis; Nutrient; Oxygen; PTPN6 gene; Parents; Parkin; Pathway interactions; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphorylation Site; Phosphotransferases; Protein Tyrosine Phosphatase; Proteins; Proteoglycan; Recombinants; Regulation; Research; Role; Second Primary Cancers; Sensitivity and Specificity; Signal Transduction; Specificity; Stromal Cells; Tamoxifen; Technology; Testing; Time; Transgenic Mice; Translating; Tumor Angiogenesis; VEGFA gene; Vascular Endothelium; Vascularization; Work; Zebrafish; adhesion receptor; aggressive breast cancer; angiogenesis; antagonist; attenuation; cancer cell; cell behavior; cellular engineering; clinically relevant; combat; confocal imaging; experimental study; genetic signature; in vivo; inducible Cre; inhibitor; innovation; interdisciplinary approach; mTOR Inhibitor; mTOR inhibition; malignant breast neoplasm; mortality; mouse model; mutant; nano-string; neoplastic cell; neovascularization; novel; novel therapeutics; nutrient deprivation; overexpression; paracrine; perlecan; predict clinical outcome; programs; promoter; receptor; sensor; transcriptome; transcriptomics; tumor; tumor microenvironment; tumor progression; tumorigenesis; tumorigenic; ultra high resolution

Chief among female mortality is the development, progression, and metastasis of aggressive breast cancer. The bidirectional communication between the neoplastic cells and the tumor microenvironment, which supplies oxygen and nutrients, is essential for promoting unchecked tumorigenic development, aberrant neovascularization, and widespread metastasis. Thus, a better understanding of embedded cues and soluble messages exchanged between these two compartments will prove invaluable for furthering our knowledge of the pathobiology of cancer and for reliably predicting clinical outcomes. Our central hypothesis is that endorepellin, a proteolytic fragment of perlecan, a multi-domain heparan sulfate proteoglycan, exploits a dual-receptor antagonism to preclude endothelial cells from participating in tumor angiogenesis. This working hypothesis is based on an impactful and promising body of work all generated under the auspices of this grant. We discovered that: [a] Endorepellin simultaneously ligates, with high affinity, VEGFR2 and the 21 integrin thereby constituting the molecular basis of “dual receptor antagonism”. Engagement of both receptors underscores the exquisite sensitivity and specificity of endorepellin in targeting the endothelia. [b] Endorepellin triggers co-internalization of VEGFR2 and 21 integrin with concurrent activation of the SHP-1 tyrosine phosphatase and attenuation of VEGFA signaling. [c] Endorepellin induces endothelial cell autophagy in a Peg3-dependent manner by modulating Beclin 1, LC3, and p62 expression, processing, and cellular localization. [d] Endorepellin evokes protracted activation of the energy-sensor kinase AMPK, irrespective of energy levels. Indeed, this regulation is considered non-canonical as AMPK phosphorylation occurred under nutrient-enriched conditions. [e] Downstream of AMPK, endorepellin evokes autophagic flux in endothelial cells that mechanistically parallels the mTOR inhibitor, Torin 1. These striking findings demonstrate that protracted and sustained autophagy is a novel mechanism by which endorepellin promotes angiostasis, independent of nutrient deprivation. Based on these discoveries, we plan to: [1] Elucidate the mechanism of endorepellin-evoked endothelial cell stress, autophagy and mitophagy. [2] Unravel the mechanism by which endorepellin induces autophagic suppression of HAS2. [3] Generate novel mouse models of tumorigenesis to explain the pro-autophagic and anti-angiogenic programs activated by endorepellin. These concerted research aims will enable us to translate our findings, procured with highly innovative mouse models of stromal autophagy, into clinically relevant paradigms. The discovery of endorepellin-induced endothelial cell autophagy downstream of dual receptor antagonism will lead to new therapeutic advances that actively induce autophagy within the tumor microenvironment to combat this devastating disease.

女性死亡的首要原因是侵袭性乳腺癌的发展、进展和转移。这个 肿瘤细胞和肿瘤微环境之间的双向通讯，为肿瘤细胞提供 氧气和营养素，是促进不受控制的肿瘤发生发展所必需的，异常 新生血管和广泛的转移。因此，更好地理解嵌入的线索和可溶性 在这两个隔间之间交换的信息将被证明是非常宝贵的，对于进一步了解 癌症的病理生物学和可靠地预测临床结果。 我们的中心假设是，内胚层蛋白是Perlecan的蛋白水解性片段，是一种多结构域硫酸肝素 蛋白多糖，利用双受体拮抗作用阻止内皮细胞参与肿瘤 血管生成。这一工作假设是基于一系列有影响力和前景的工作，这些工作都是在 这笔赠款的赞助人。我们发现：[A]内毒素以高亲和力同时连接， VEGFR2和-2--1整合素构成了“双受体拮抗”的分子基础。 这两种受体的结合突显了内层促性腺激素在靶向方面的高度敏感性和特异性 内皮细胞。[B]内排斥素触发VEGFR2和21整合素的共同内化 SHP-1酪氨酸磷酸酶的激活和VEGFA信号的减弱。[C]内排斥素诱导 通过调节Beclin 1、Lc3和p62的表达，以Peg3依赖的方式实现内皮细胞自噬。 加工和细胞定位。[D]内排斥素引起能量感受器激酶的持久激活 AMPK，无论能量水平如何。事实上，这一规定被认为与AMPK一样不规范 磷酸化发生在营养丰富的条件下。[E]AMPK下游，内胚层蛋白激活 内皮细胞的自噬通量在机械上与mTOR抑制剂Torin 1平行。这些惊人的 研究结果表明，持久和持续的自噬是内皮化的一种新机制 促进血管稳定，不受营养剥夺的影响。基于这些发现，我们计划： [1]阐明内质网促内皮细胞应激、自噬和有丝分裂吞噬的机制。 [2]解开内脂蛋白诱导HAS2自噬抑制的机制。 [3]建立新的小鼠肿瘤发生模型来解释促自噬和抗血管生成程序 被内胚层蛋白激活。 这些协调一致的研究目标将使我们能够翻译我们的发现，通过高度创新的鼠标获得 间质自噬的模型，转化为临床相关的范例。内源性促性腺激素的发现 双受体拮抗剂下游的内皮细胞自噬将带来新的治疗进展 在肿瘤微环境中积极诱导自噬，以对抗这种毁灭性的疾病。